Applications of cell cultures in research MCQs With Answer are essential for B. Pharm students studying pharmacology, toxicology, and biopharmaceutical development. Cell culture applications enable in vitro models for drug screening, ADME-Tox studies, vaccine and monoclonal antibody production, tissue engineering, and mechanistic pharmacological research. Understanding cell-based assays, primary versus continuous cell lines, sterile technique, media formulation, and contamination control prepares students for practical lab work and research design. This topic links theory to real-world drug discovery, quality control, and regulatory considerations in pharmaceutical development. Now let’s test your knowledge with 50 MCQs on this topic.
Q1. What is the primary advantage of using cell cultures in early-stage drug screening?
- Eliminates the need for any animal or human testing
- Provides controlled in vitro conditions for mechanistic and high-throughput screening
- Guarantees clinical efficacy in humans
- Is universally cheaper than all alternative methods
Correct Answer: Provides controlled in vitro conditions for mechanistic and high-throughput screening
Q2. Which statement best distinguishes primary cell cultures from continuous (immortalized) cell lines?
- Primary cells are easier to culture indefinitely
- Continuous cell lines maintain original tissue phenotype longer than primary cells
- Primary cells are directly isolated from tissues and have limited lifespan whereas continuous lines proliferate indefinitely
- Continuous lines cannot be used in drug metabolism studies
Correct Answer: Primary cells are directly isolated from tissues and have limited lifespan whereas continuous lines proliferate indefinitely
Q3. Which of the following is a common method to assess cell viability in culture?
- Gram staining
- MTT or resazurin assay
- PCR amplification of housekeeping gene
- ELISA for secreted cytokines only
Correct Answer: MTT or resazurin assay
Q4. Which media component is critical for supplying amino acids, vitamins, and energy substrates to cultured cells?
- Antibiotics
- Basal culture medium (e.g., DMEM, RPMI)
- Trypsin
- Detergent
Correct Answer: Basal culture medium (e.g., DMEM, RPMI)
Q5. What is the main function of fetal bovine serum (FBS) in cell culture media?
- Act as a selective antibiotic
- Supply growth factors, hormones, and attachment factors
- Fix cells for microscopy
- Prevent cell adhesion
Correct Answer: Supply growth factors, hormones, and attachment factors
Q6. Which technique is used to detach adherent cells from culture flasks for passaging?
- PCR
- Trypsinization or enzymatic dissociation
- Gram staining
- Lyophilization
Correct Answer: Trypsinization or enzymatic dissociation
Q7. What does “confluency” refer to in adherent cell cultures?
- The percentage of cells actively dividing
- The proportion of the culture surface covered by cells
- The concentration of antibiotics in media
- The number of passages a cell line has undergone
Correct Answer: The proportion of the culture surface covered by cells
Q8. Which contamination is most insidious and often requires PCR or DNA staining for detection?
- Fungal spores
- Bacterial contamination visible by turbidity
- Mycoplasma contamination
- Yeast contamination with hyphae
Correct Answer: Mycoplasma contamination
Q9. For sterile technique in cell culture, which practice is essential?
- Working in a Class II biosafety cabinet and using aseptic technique
- Opening culture flasks near an open window
- Using non-sterile gloves to handle reagents
- Heating media on an open flame
Correct Answer: Working in a Class II biosafety cabinet and using aseptic technique
Q10. What is the purpose of cryopreservation of cell lines?
- Increase mutation rates intentionally
- Store cells long-term at very low temperatures to preserve viability and genetics
- Differentiate stem cells into specific lineages
- Eliminate the need for a growth medium
Correct Answer: Store cells long-term at very low temperatures to preserve viability and genetics
Q11. What is a spheroid in 3D cell culture models?
- A monolayer of cells on a petri dish
- A 3D aggregate of cells that better mimics in vivo tissue architecture
- A type of bacterial colony contaminant
- A freeze-dried cell pellet
Correct Answer: A 3D aggregate of cells that better mimics in vivo tissue architecture
Q12. Which assay commonly measures cytotoxicity via metabolic activity conversion of tetrazolium salts?
- Western blot
- MTT assay
- Flow cytometry scatter plot
- Chromatography
Correct Answer: MTT assay
Q13. Why are cell authentication and STR profiling important in cell culture research?
- To increase growth rate of cells
- To confirm cell line identity and prevent misidentification or cross-contamination
- To sterilize cells before use
- To measure media pH
Correct Answer: To confirm cell line identity and prevent misidentification or cross-contamination
Q14. In ADME-Tox studies, which cell system is frequently used to assess hepatic metabolism?
- Neuronal primary cultures
- Hepatocyte cultures or HepG2 cell line
- Keratinocyte monolayers
- Cardiomyocyte spheroids only
Correct Answer: Hepatocyte cultures or HepG2 cell line
Q15. What is co-culture in cell biology?
- Growing cells under antibiotic-free conditions
- Culturing two or more different cell types together to study interactions
- Keeping cells in separate flasks with same medium
- Freezing cells with serum
Correct Answer: Culturing two or more different cell types together to study interactions
Q16. Which factor is essential when scaling up cell culture for biopharmaceutical production?
- Ignoring dissolved oxygen and pH
- Bioreactor design, control of shear, pH, oxygen, and nutrient supply
- Using only T-flasks regardless of volume
- Avoiding any form of cell monitoring
Correct Answer: Bioreactor design, control of shear, pH, oxygen, and nutrient supply
Q17. What role do feeder layers play in some primary or stem cell cultures?
- Act as antibiotics to prevent contamination
- Provide growth-supporting factors and substrate for attachment
- Function as cryoprotectants
- Cause immediate differentiation of cells
Correct Answer: Provide growth-supporting factors and substrate for attachment
Q18. Which of the following is a key regulatory consideration for cell-based assays in drug development?
- Standardization, validation, and documentation under GLP/GCCP
- Using unvalidated homemade reagents only
- Never reporting passage number
- Deliberately contaminating cultures to test robustness
Correct Answer: Standardization, validation, and documentation under GLP/GCCP
Q19. What does “serum-free media” offer for certain cell culture applications?
- Increased variability and unknown factors
- Defined composition that reduces variability and is useful for mechanistic studies
- Guaranteed immortalization of primary cells
- Prevention of all forms of contamination
Correct Answer: Defined composition that reduces variability and is useful for mechanistic studies
Q20. Which imaging technique is commonly used to assess cell morphology and confluency in culture?
- Transmission electron microscopy for routine checks
- Phase-contrast light microscopy
- Mass spectrometry
- Nuclear magnetic resonance (NMR)
Correct Answer: Phase-contrast light microscopy
Q21. What is mycoplasma’s impact on cell culture experiments?
- No impact—mycoplasma is harmless
- It can alter cell metabolism, growth, and experimental results without visible turbidity
- It only affects bacterial cultures, not eukaryotic cells
- It always produces visible fungal structures
Correct Answer: It can alter cell metabolism, growth, and experimental results without visible turbidity
Q22. Which cell-based assay is best suited to measure apoptosis specifically?
- Bacterial culture turbidity
- Annexin V/propidium iodide staining with flow cytometry
- pH meter reading
- Gram staining
Correct Answer: Annexin V/propidium iodide staining with flow cytometry
Q23. In transfection experiments, what is the primary purpose of using a transfection reagent or electroporation?
- To remove cells from culture
- To introduce nucleic acids (DNA/RNA) into cells for gene expression or silencing
- To change cell morphology permanently
- To sterilize the culture
Correct Answer: To introduce nucleic acids (DNA/RNA) into cells for gene expression or silencing
Q24. What is an organoid?
- A bacterial biofilm
- A miniaturized, self-organized 3D tissue model derived from stem cells that resembles an organ
- A frozen cell pellet used for cryopreservation
- An antibiotic-resistant cell line
Correct Answer: A miniaturized, self-organized 3D tissue model derived from stem cells that resembles an organ
Q25. Which parameter is commonly monitored online in bioreactors to maintain optimal cell growth?
- Presence of color in media
- pH, dissolved oxygen (DO), and temperature
- Weight of the flask lid
- Number of pipette tips used
Correct Answer: pH, dissolved oxygen (DO), and temperature
Q26. Why is antibiotic overuse in routine cell culture discouraged?
- It guarantees sterile cultures forever
- It masks low-level contamination, can induce resistance, and may affect cell physiology
- It increases growth rates of all cells
- It replaces the need for aseptic technique
Correct Answer: It masks low-level contamination, can induce resistance, and may affect cell physiology
Q27. Which cell line is classically used as a model for neuronal studies?
- HeLa
- PC12 or SH-SY5Y
- CHO
- E. coli
Correct Answer: PC12 or SH-SY5Y
Q28. In toxicity testing, what advantage do human cell lines offer over animal models?
- They are always cheaper than animal tests
- They can provide human-specific metabolic or receptor responses and reduce interspecies differences
- They remove the need for any further validation in vivo
- They never show variability
Correct Answer: They can provide human-specific metabolic or receptor responses and reduce interspecies differences
Q29. Which preservation agent is commonly used as a cryoprotectant during freezing?
- Formaldehyde
- Dimethyl sulfoxide (DMSO)
- Sodium chloride
- Phenol red
Correct Answer: Dimethyl sulfoxide (DMSO)
Q30. What does the term “passage number” indicate?
- The number of times cells have been fed with media
- The number of subcultures a cell population has undergone since isolation
- The number of cells counted in a hemocytometer
- The number of different media types tried
Correct Answer: The number of subcultures a cell population has undergone since isolation
Q31. Which readout is commonly used in cell-based reporter assays?
- Reporter gene expression such as luciferase or GFP
- Visual turbidity of medium
- Cell pellet weight only
- pH of culture media exclusively
Correct Answer: Reporter gene expression such as luciferase or GFP
Q32. How do adherent and suspension cell cultures fundamentally differ?
- Adherent cells float; suspension cells attach to surfaces
- Adherent cells require a surface for attachment; suspension cells grow freely in medium
- Only adherent cells can be cultured in bioreactors
- Suspension cultures do not require sterile technique
Correct Answer: Adherent cells require a surface for attachment; suspension cells grow freely in medium
Q33. What is Good Cell Culture Practice (GCCP) primarily focused on?
- Ensuring reproducibility, traceability, biosafety, and ethical conduct in cell culture work
- Maximizing contamination rates for stress testing
- Permanent use of antibiotics in all experiments
- Avoiding documentation
Correct Answer: Ensuring reproducibility, traceability, biosafety, and ethical conduct in cell culture work
Q34. Which technique is preferred to detect and quantify mycoplasma contamination?
- Gram staining
- PCR-based assays or specific fluorescent staining
- Routine light microscopy only
- Plating on LB agar
Correct Answer: PCR-based assays or specific fluorescent staining
Q35. Why are human induced pluripotent stem cells (iPSCs) valuable in pharmaceutical research?
- They are terminally differentiated and cannot be manipulated
- They provide patient-specific models for disease modeling, toxicity testing, and regenerative medicine
- They replace the need for any clinical trials
- They are always cheaper than primary cells
Correct Answer: They provide patient-specific models for disease modeling, toxicity testing, and regenerative medicine
Q36. Which assay can quantify secreted proteins such as cytokines from cultured cells?
- ELISA
- Gram staining
- Hemocytometer counting
- Phase-contrast microscopy
Correct Answer: ELISA
Q37. What is the significance of multiplicity of infection (MOI) in viral transduction of cells?
- It measures the concentration of antibiotics
- It indicates the ratio of infectious viral particles to target cells and influences transduction efficiency
- It refers to the number of passages
- It is unrelated to viral gene delivery
Correct Answer: It indicates the ratio of infectious viral particles to target cells and influences transduction efficiency
Q38. Which factor most strongly influences differentiation of stem cells in culture?
- Ambient room color
- Microenvironmental cues such as growth factors, substrate stiffness, and cell–cell interactions
- The brand of flask used only
- The presence of antibiotics exclusively
Correct Answer: Microenvironmental cues such as growth factors, substrate stiffness, and cell–cell interactions
Q39. In drug permeability studies, which cell model mimics intestinal epithelium?
- CHO cells
- Caco-2 cell monolayer
- HepG2 hepatocytes
- HeLa cancer cells
Correct Answer: Caco-2 cell monolayer
Q40. What is the role of trypsin-EDTA in cell culture handling?
- To stain cells for microscopy
- To enzymatically detach adherent cells by breaking cell–substrate and cell–cell interactions
- To serve as a cryoprotectant
- To act as a nutrient supplement
Correct Answer: To enzymatically detach adherent cells by breaking cell–substrate and cell–cell interactions
Q41. Which parameter is important to record for reproducibility in cell-based experiments?
- Passage number, seeding density, medium composition, and incubation conditions
- Only the time of day experiments were done
- Only the brand of gloves used
- Ambient music played in lab
Correct Answer: Passage number, seeding density, medium composition, and incubation conditions
Q42. How do 3D cultures improve the physiological relevance of in vitro studies?
- They are easier to image than 2D cultures
- They provide cell–cell and cell–matrix interactions, gradients of nutrients and drugs, and more in vivo-like responses
- They always remove the need for biochemical assays
- They eliminate any variability in experiments
Correct Answer: They provide cell–cell and cell–matrix interactions, gradients of nutrients and drugs, and more in vivo-like responses
Q43. What is a common quality control step before using a thawed cell vial for experiments?
- Immediate use without recovery
- Allow cells to recover, assess viability, morphology, and mycoplasma status
- Heat the vial to 100°C
- Discard cells after thawing
Correct Answer: Allow cells to recover, assess viability, morphology, and mycoplasma status
Q44. Which cellular feature is often monitored to indicate differentiation in muscle cell cultures?
- Formation of multinucleated myotubes and expression of muscle-specific markers
- Decrease in medium color only
- Production of bacterial colonies
- Spontaneous detachment of all cells
Correct Answer: Formation of multinucleated myotubes and expression of muscle-specific markers
Q45. What is the purpose of a cell bank in pharmaceutical research?
- To store documentation only
- To maintain master and working stocks of authenticated, quality-controlled cell lines for reproducible use
- To grow bacteria for antibiotic production
- To hold expired reagents
Correct Answer: To maintain master and working stocks of authenticated, quality-controlled cell lines for reproducible use
Q46. Which of the following best describes an endpoint assay?
- An assay that continuously measures cells in real time
- An assay measured after a fixed incubation period to quantify a final outcome such as viability or protein level
- A technique to freeze cells
- A method of sterile filtration
Correct Answer: An assay measured after a fixed incubation period to quantify a final outcome such as viability or protein level
Q47. In cell-based assays for receptor pharmacology, what does EC50 represent?
- Concentration of compound producing 50% of maximal effect
- Total cell count at 50 hours
- Number of experiments repeated 50 times
- Evaporation coefficient
Correct Answer: Concentration of compound producing 50% of maximal effect
Q48. Which method is best to quantify cell proliferation over several days in a microplate format?
- Single-point Gram staining
- BrdU incorporation or cell counting assays (e.g., automated cell counters, crystal violet staining)
- pH strip testing only
- Using a thermometer
Correct Answer: BrdU incorporation or cell counting assays (e.g., automated cell counters, crystal violet staining)
Q49. What is the major limitation of 2D monolayer cultures for drug testing?
- They perfectly mimic whole-organ responses
- Lack of 3D architecture, gradients, and multicellular interactions present in tissues which can affect drug response
- They are incompatible with most assays
- They never require quality control
Correct Answer: Lack of 3D architecture, gradients, and multicellular interactions present in tissues which can affect drug response
Q50. Why is documentation of cell line provenance important in pharmaceutical studies?
- Provenance is optional and rarely useful
- To ensure traceability, legal compliance, reproducibility, and to prevent use of misidentified or contaminated lines
- To avoid using any quality control tests
- To reduce the cost of experiments by skipping authentication
Correct Answer: To ensure traceability, legal compliance, reproducibility, and to prevent use of misidentified or contaminated lines

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